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A clsm scans objects point by point by a finely focussed laser beam. The fluorescence light which is emitted by the specimen is detected by photomultipliers and is used to modulate the light intensity of a display monitor which is synchronized to the laser scanning motion. The image of the specimen is then formed on a display monitor. The advantages of this optical scanning method are that less stray light is produced and that in addition to various optical contrasting methods brightness and contrast of the image can be varied electronically.
The total energy applied to the specimen during a scan is much lower than conventional fluorescence microscopy. Furthermore, well-defined thin optical sections through the specimen can be reassembled to 3D-reconstructions. Due to the high intensity of the laser beam and the fine resolution of the fluorescence image further possibilities arise. The confocal system allows the observation over longer periods of time in bright field and fluorescence mode.
Our microscopy facility, integral part of the Faculty of Life Sciences, comprises advanced video, light and electron microscopy combined with atomic element analysis, as well as means for documentation of biological topics by photography and scientific videography.
In our research, we elucidate structures of various degrees of complexity which will thereby contribute to a better understanding of function, ontogenetic and physiologic processes, as well as phylogenetic relationships. This includes widespread cooperation with other research institutions in Austria and abroad.
We teach and train students in light-and electron microscopic techniques and we supervise Bachelor and Master Students with a sound interest in structural/ functional relationship.
We assist our customers with technical equipment and expert guidance in order for them to obtain the maximum benefit to execute their research projects. Besides of members of faculty and the university, we welcome customers from other educational and research institutions and from industry.
Methods & Expertise for Research Infrastructure
In fluorescence microscopy cellular structures are stained by fluorescent dyes; they are excited by certain wavelengths and emit fluorescence light of longer wavelength. Specific filter sets allow for observation of the emitted light on a dark background.
Staining can be used for:
•visualization of sub-resolution structures by organelle-specific dyes
•identification of visible and sub-resolution structures by organelle-specific dyes
•immuno-labelling of proteins
•fluorescence-in-situ-hybridization of DNA (FISH)
•visualization of proteins in living cells by green fluorescent protein technique (GFP)
•quantitative analysis of cell content
•measuring of physiological parameters such as membrane potential, yield of photosynthesis or respiration
In addition to fluorescence staining, quite a few biological structures emit fluorescent light naturally upon excitation. A well known example is chlorophyll which can be excited by different wavelenghts and emits red light. This phenomenon is called autofluorescence.
For further information about the principles and techniques of fluorescence microscopy, please visit our electronic script about light microscopy:
Core Facility Cell Imaging and Ultrastructure Research
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